ELISA Kit for Angiotensin II (AngII)

Ang-II; Angiotensin-2

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiotensin II (AngII).
No significant cross-reactivity or interference between Angiotensin II (AngII) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiotensin II (AngII) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiotensin II (AngII) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-91 87
EDTA plasma(n=5) 96-103 99
heparin plasma(n=5) 90-104 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensin II (AngII) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensin II (AngII) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Angiotensin II (AngII) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-101% 82-94% 84-99% 97-105%
EDTA plasma(n=5) 96-105% 93-101% 85-94% 96-103%
heparin plasma(n=5) 79-95% 87-94% 85-94% 89-96%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Molecular medicine Angiotensin-Converting Enzyme 2 Overexpression Remarkably Ameliorated Glomerular Injury in a Rat Model of Diabetic Nephropathy: A Comparison with ACE Inhibition MolMed: 10_11_liu
The European Journal of Clinical Investigation Mesangial medium with IgA1 from IgA nephropathy inhibits nephrin expression in mouse podocytes Wiley: source
Immunopharmacology and Inflammation Evidence for the complementary and synergistic effects of the three-alkaloid combination regimen containing berberine, hypaconitine and skimmianine on the ulcerative colitis rats induced by trinitrobenzene-sulfonic acid ScienceDirect: S0188440910001049
Nephrology Effect of aggregated immunoglobulin A1 from immunoglobulin A nephropathy patients on nephrin expression in podocytes Wiley: source
PLoS One Angiotensin II and Angiotensin-(1-7) in Paraventricular Nucleus Modulate Cardiac Sympathetic Afferent Reflex in Renovascular Hypertensive Rats PubMed: PMC3527547
BioMed Research International Heterogeneous Downregulation of Angiotensin II AT1-A and AT1-B Receptors in Arterioles in STZ-Induced Diabetic Rat Kidneys Hindawi: 947506
34 Biglcan is beneficial in angiotensin II induced heart failure by preventing cardiac inflammation and remodeling improving LV function and mortality by preventing transdifferentiation of myofibroblasts Oxfordjournals: Source
Pflügers Archiv-European Journal of Physiology SOD1 gene transfer into paraventricular nucleus attenuates hypertension and sympathetic activity in spontaneously hypertensive rats Pubmed: 23114721
Eur J Pharmacol. Angiotensin II and vasopressin are involved in the defense system against anaphylactic hypotension in anesthetized rats Pubmed:24650734
J Mol Cell Cardiol. Increased LDL electronegativity in chronic kidney disease disrupts calcium homeostasis resulting in cardiac dysfunction Pubmed:25871829
Mol Med Rep. High glucose induces activation of the local renin?angiotensin system in glomerular endothelial cells Pubmed:24337709
Mol Med Rep. Angiotensin II promotes the progression of human gastric cancer Pubmed:24424956
PLOS ONE Intermedin in the Paraventricular Nucleus Attenuates Cardiac Sympathetic Afferent Reflex in Chronic Heart Failure Rats Pubmed:24709972
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Biochim Biophys Acta. Cathepsin G activity lowers plasma LDL and reduces atherosclerosis Pubmed:25092171
Acta Physiologica Superoxide anions in the paraventricular nucleus mediate cardiac sympathetic afferent reflex in insulin resistance rats Pubmed:25307720
Neuroscience Letters Administration of exogenous 1,25(OH)2D3 normalizes overactivation of the central renin-angiotensin system in 1α(OH)ase knockout mice Pubmed:25576706
34 A background Ca2+ entry pathway mediated by TRPC1/TRPC4 is critical for development of pathological cardiac remodelling PubMed: 26069213
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Nutrition Effects of Rubus occidentalis extract on blood pressure in subjects with prehypertension: randomized, double-blinded, placebo controlled clinical trial PubMed: 26740254
J Am Soc Hypertens Angiotensin-(1–7) enhances the effects of angiotensin II on the cardiac sympathetic afferent reflex and sympathetic activity in rostral ventrolateral medulla in renovascular hypertensive rats PubMed: 26428223
PLoS One. Up ReCavia (Guinea pig )lation of cystathione γ lyase and Hydrogen Sulphide in the Myocardium Inhibits the Progression of Isoproterenol–Caffeine Induced Left Ventricular Hypertrophy in Wistar Kyoto Rats Pubmed:26963622
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PLoS One. Paradoxical Sleep Deprivation Causes Cardiac Dysfunction and the Impairment Is Attenuated by Resistance Training. pubmed:27880816
Oxidative Medicine and Cellular Longevity Epigallocatechin-3-gallate Attenuates Renal Damage by Suppressing Oxidative Stress in Diabetic db/db Mice pubmed:27698952
Journal of the Renin-Angiotensin-Aldosterone System Angiotensin type 2 receptor null mice express reduced levels of renal angiotensin converting enzyme-2/angiotensin (1-7)/Mas receptor and exhibit greater high-fat diet-induced kidney injury 1470320316661871.abstract
Microvascular Research Pleiotropic effects of survivin in vascular endothelial cells. pubmed:27373518
PLOS ONE Reduced Expression of the Extracellular Calcium-Sensing Receptor (CaSR) Is Associated withActivation of the Renin-Angiotensin System (RAS) to Promote Vascular Remodeling in the Pathogenesis of Essential Hypertension. pubmed:27391973
journal of pharmacological sciences β-Aminoisobutyric acid ameliorates the renal fibrosis in mouse obstructed kidneys via inhibitionof renal fibroblast activation and fibrosis. pubmed:28433566
Experimental Animals Ginsenoside Rb1 improves cardiac function and remodeling in heart failure. pubmed:28367863
Oncotarget Effects of renal denervation on monocrotaline induced pulmonary remodeling pubmed:28187460
The American Journal of Pathology The Renin-Angiotensin System Regulates Neurodegeneration in a Mouse Model of Optic Neuritis pubmed:28919108
PLoS One Enhanced heart failure, mortality and renin activation in female mice with experimental dilated cardiomyopathy pubmed:29240788
Frontiers in Physiology Hydrogen Sulfide Attenuates Atherosclerosis in a Partially Ligated Carotid Artery Mouse model via Regulating Angiotensin Converting Enzyme 2 Expression pubmed:29066981
Neurotherapeutics Brain Renin–Angiotensin System Blockade Attenuates Methamphetamine-Induced Hyperlocomotion and Neurotoxicity Pubmed:29464572
International Journal of Cardiology ALDH2 protects against alcoholic cardiomyopathy through a mechanism involving the p38 MAPK/CREB pathway and local renin-angiotensin system inhibition in … Pubmed:29506687
Neuroscience Angiotensin II Type 1a Receptors in the Subfornical Organ Modulate Neuroinflammation in the Hypothalamic Paraventricular Nucleus in Heart Failure Rats Pubmed:29684507
American Journal of Physiology-Renal Physiology Role of (Pro) Renin Receptor in Albumin Overload-Induced Nephropathy in Rats Pubmed:29846109
Journal of the American Society of Hypertension Mesenchymal stem cell–derived microvesicles alleviate pulmonary arterial hypertension by regulating renin-angiotensin system Pubmed:29752040
Evidence-based Complementary and Alternative Medicine  Acute Biochemical, Cardiovascular, and Autonomic Response to Hyperbaric (4 atm) Exposure in Healthy Subjects Pubmed:29977313
Brazilian Journal of Medical and Biological Research Renovascular hypertension increases serum TNF and CX3CL1 in experimental Trypanosoma cruzi infection Pubmed:29590257
American Journal of Physiology-Heart and Circulatory Physiology Sex Differences in the Central and Peripheral Manifestations of Ischemia-induced Heart Failure in Rats Pubmed: 30289294
Toxicology Mechanisms and Methods Long-term exposure to crotonaldehyde causes heart and kidney dysfunction through induction of inflammatory and oxidative damage in male Wistar rats Pubmed: 30426860
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International Journal of Ophthalmology Serum microRNA-221 as a biomarker for diabetic retinopathy in patients associated with type 2 diabetes Pubmed: 30588418
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The American Journal of the Medical Sciences T lymphocyte-based renin angiotensin system in obesity
Journal of Alzheimers Disease Cerebrospinal Fluid Changes in the Renin-Angiotensin System in Alzheimer's Disease Pubmed: 31594235
Andrologia Seminal levels of angiotensin II and angiotensin II type 2 receptor expression on spermatozoa in varicocele patients: Relation to fertility status Pubmed: 31989676
Molecular Medicine Reports BPA disrupts 17‑estradiol‑mediated hepatic protection against ischemia/reperfusion injury in rat liver by upregulating the Ang II/AT1R signaling pathway Pubmed: 32319667
MOLECULAR MEDICINE REPORTS The sympathetic transmitter norepinephrine inhibits VSMC proliferation induced by TGFβ by suppressing the expression of the TGFβ receptor ALK5 in aorta … Pubmed: 32319652
Cell Mol Neurobiol Predator Scent-Induced Sensitization of Hypertension and Anxiety-like Behaviors Pubmed: 33201417
biochemistry biophysical reports Effect of repetitive potassium iodide on thyroid and cardiovascular functions in elderly rats Pubmed: 33024842
bioscience reports Angiotensin II enhances the proliferation of Natural Killer/T-cell lymphoma cells via activating PI3K/Akt signaling pathway Pubmed: 32969473
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY A (pro) renin receptor decoy peptide PRO20 protects against adriamycin-induced nephropathy by targeting intrarenal renin-angiotensin system Pubmed: 32865014
Acta Endocrinologica-Bucharest THE ROLE OF HIGH FOLLICULAR LEVELS OF ANGIOTENSIN II AND VASCULAR ENDOTHELIAL GROWTH FACTOR IN ANTICIPATING THE … Pubmed: 32685035
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